• KSBB Journal
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• v.5 no.1
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• pp.49-58
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• 1990

We constructed hybrid plasmids to allow controlled and extracellular production of human alpha-interferon in Escherichia coli. The hybrid plassmids were constructed by transferring alpha-lFN gene from plasmid Hif-2h which has the alpha-lFN gene at PstI restriction site of pBR322, to plasmids pIN -IIIB3 and pIN-IIIC3 at restriction sites between HindIII and BamHI. Plasmids pIN-IIIB3 and pIN-IIIC3 carry E. coli lipoprotein promoter, lac promoter and operator in tandem. The plasmids also have lacl genes which encode for lac repressors, which allows controlled expression of genes cloned to the plasmids by using of inducer IPTG. Lipoprotein signal sequence is located just ahead of cloning sites of the plasmids, which helps cells to excrete or secrete cloned gene products. Plasmid pUC9 was used as a intermediate vector for transferring of alpha-lFN gene from Hif-2h to pIN vectors in order to solve the problem of different restriction sites between Hif-2h and pIN vectors.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.418.1-418.1
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• 2002

Recombinant interferon alpha is widely used for the treatment of diseases including chronic hepatitis C. However. it has drawbacks such as relatively short serum half-life and rapid clearance like other therapeutic proteins. Using PFGylFltioo which is one of the well-established methods to fulfill the requirements of a long-lasting form of protein. wIe prepared mono-PEG modified interferon alpha-2a in which polyethylene glycol moiety was covalently attached to the amino groups of interfelon alpha-2a. (omitted)

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.166-172
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• 2000

Prior to a clinical trial, the in vitro and in vivo antitumor effects of a new recombinant human interferon ${\alpha}-2a$ (rHu/IFN ${\alpha}-2a$) with/without hydroxyurea (HU) were investigated using chronic myelogenous leukemia (CML)-derived cell lines (K562 and KU812F) and BALB/c nude. mice transplanted with KU812F cells. The rHu/IFN ${\alpha}-2a$ ($10^4-10^6IU/ml$) strongly inhibited proliferation of both cell lines and the combined treatments with HU ($10{\mu}g/ml$) were more effective. In nude mice transplanted with KU812F cells. rHu/IFN ${\alpha}-2a(1{\times}10^6IU$) inhibited tumor growth by 42-65% at 15-21 days post-transplantation (DPT). The combined treatment of rHu/IFN ${\alpha}-2a (5{\times}10^5IU$) with HU (0.25mg/g b.w.) inhibited the tumor growth by 48-67% at 12-21 DPT. In addition, the treatment of rHu/IFN ${\alpha}-2a$ ($5{\times}10^6IU\;or\;1{\times}10^7IU$) rejected tumor transplantation by 40%. These results suggest that the new rHU/IFN ${\alpha}-2a$ alone or with HU is effective on CML cell lines.

• The Journal of Internal Korean Medicine
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• v.20 no.1
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• pp.143-152
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• 1999

Dansam, the root of Salvia miltiorrhiza Bge, (Labiatae), has a bitter taste and a slightly 'cold' property, and is nontoxic. In the present study, effect of Dansam on nitric oxide (NO) generation from peritoneal macrophags was examined. Dansam had no effect on NO generation by itself, whereas recombinant interferon-${\gamma}\;(rIFN-{\gamma})$ alone had modest activity. When Dansam was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO generation in a dose-dependent manner, The optimal effect of Dansam on NO generation was shown at 6 hr after treatment with $rIFN-{\gamma}$. Furthermore, the effect of Dansam was mainly dependent on Dansam-induced tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ secretion. These results suggest that Dansam induces NO generation from macrophages by the result of Dansam-induced $TNF-{\alpha}$ secretion.

• Journal of Periodontal and Implant Science
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• v.24 no.1
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• pp.155-164
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• 1994

치조골흡수는 만성치주질환의 전형적인 증상이다. 골흡수에 작용하는 여러 요인들 중에서도, 특히 최근에 들어서 몇몇 cytokine들에 대한 관심이 높아지고 있는데, interleukin-1(IL-1), tumor necrosis factor(TNF) 및 interleukin-6(IL-6) 등이 치주질환의 진행과정에서 중요한 치조골흡수요인으로 제안되고 있다. 본 연구의 목적은 신생쥐의 골조직 배양실험을 통해서 recombinant human $interleukin-1{\beta}$ ($rHuIL-1{\beta}$), recombinant human tumor necrosis $factor-{\alpha}$($rHuTNF-{\alpha}$) 및 recombinant human interleukin-6(rHuIL-6) 의 골흡수 유도효과를 알아보고, cyclooxygenase 억제제인 indomethacin과 recombinant murine $interferon-{\gamma}$($rMurIFN-{\gamma}$)가 이들 cytokine의 골흡수 유도능력에 미치는 영향을 알아봄으로써 이들 cytokine의 작용기구에 대해서 알아보고자 하는데 있다. 생후 1-2일된 쥐에게 $1{\mu}Ci^{45}CaCl_2$를 피하주사하고 4일 후에 쥐를 희생시켜 $^{45}Ca$ 로 표지된 두개골을 얻어 24시간 전배양 후, 각 cytokine ($rHuIL-1{\beta}$, $rHuTNF-{\alpha}$ 및 rHuIL-6)과 cytokine 및 첨가약제 (indomethacin 및 $rMurIFN-{\gamma}$)가 함유된 배지로 교환하여 48시간 배양한다. 골흡수 유도효과는 두개골에서 48시간의 배양 중 유리되는 $^{45}Ca$의 방사능 정도로 평가하였다. 본 연구를 통해 다음과 같은 결과를 얻었다. 1. $rHuIL-1{\beta}$ ($10^{-12}-10^{-9}M$) 및 $rHuTNF-{\alpha}$ ($10^{-10}-10^{-8}M$)는 농도변화에 따르는 골흡수 유도효과를 보였으나 , rHuIL-6 ($10^{-10}-10^{-8}M$)는 유의할 만한 효과를 보이지 않았다. 2. Indomethacin ($10^{-6}M$)은 $rHuIL-1{\beta}$ 및 $rHuTNF-{\alpha}$의 골흡수 유도작용에 유의할 만한 억제효과를 나타내지 않았다. 3. $rMurIFN-{\gamma}$ (1000 U/ml) 은 $rHuIL-1{\beta}$ 및 $rHuTNF-{\alpha}$의 골흡수 유도작용에 유의한 억제효과를 나타내었다. 본연구를 통해 치주질환 환자의 치주조직에서 검출되는 $IL-1{\beta}$ 및 $TNF-{\alpha}$가 치조골 흡수에 중요한 역할을 할 것으로 생각된다.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.1 no.1
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• pp.79-89
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• 1998

Purpose: Though many antiviral or immunomodulatory agents have been used in patients with chronic HBV hepatitis, interferon is considered to be the only effective therapeutic agent so far. Among immunomodulatory agents, thymodulin, the oral form of thymosin, is currently in clinical trial. We compared the efficacy of alfa-interferon therapy alone with a combined therapy of alfa-interferon and thymodulin in children with chronic active hepatitis B. Method: Twenty three children aged 4.4~13.7 years who were known to be positive for HBsAg and HBeAg in serum for at least 6 months and who had biopsy-proven chronic active hepatitis were given either combined therapy of alfa-interferon and thymodulin or alfa-interferon alone, and all children were HBV DNA positive in their serum at the beginning. Follow-ups have been done for at least 1 year after a 6 month course of therapy and clearance of viral replication markers has been evaluated. Results: 1) During follow up period, 11 (48%) children were seroconverted to anti-HBe and were cleared of HBV DNA from their serum. However, 2 of them relapsed after discontinuance of interferon therapy. 2) Seroconversion occurred more frequently among those who had not been vertically transmitted, had elevated serum ALT levels and low HBV DNA levels before interferon therapy. 3) There was no significant advantage of the combined therapy with thymodulin compared to interferon therapy alone. Conclusion: Combined therapy of alfa-interferon and thymodulin failed to demonstrate synergistic effect. We think that combination therapies of alfa-interferon with other antiviral or immunomodulatory agents need to be studied in order to achieve better therapeutic responses.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.4
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• pp.301-305
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• 2001

The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

• YAKHAK HOEJI
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• v.39 no.6
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• pp.622-630
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• 1995

Inoculation of Friend anemia virus, which was a Kind of retro virus such as HIV, results splenomegaly, anemia, the increase of WBC counts and reverse transcriptase activity in serum. These results were due to the inhibition of the differentiation of erythroid progenitor cell by the FVA at the spleen. Using these as index of antiviral effects, we pursued the establishment of in vivo screening method for the new anti-ADS drugs. Among zidovudine, didanosine and zalcitabine, which were already approved as anti-AIDS drugs, treatment of zidovudine for 18 days in BALB/c mice inoculated with Friend anemia virus resulted the most potent inhibitory effects on the splenomegaly, the increase of WBC counts and reverse transcriptase activity, but did not recovered the anemia due to the tomcity of zidovudhie itself on the bone marrow. The antiviral effects of zidovudine was reduced in case of zidovudine treatment 7 days after Friend anemia virus inoculation. These results suggested that the sooner treatment of zidovudine would be better improved when the virus was inoculated. Human recombinant interferon itself .alpha. did not showed the antiviral activity against Friend anemia virus and did not also affected the antiviral activity of zidovudine. These results suggested that Friend anemia virus would be used as a tool in vivo screening method for the Lobster of reverse transcriptase.

• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.365-375
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• 1995

The anemia-inducing strain of Friend virus (FVA) is a murine retrovirus which stimulates the proliferation of erythroid progenitor cells. The progenitor cells synthesized by FVA-stimulation are unable to proceed with differentiation and accumulate in the spleen resulting in splenomegaly in infected mice. Using FVA-inoculated mice as a model, we have investigated the antiretroviral effects of 2',3'-dideoxycytidine (ddC) and recombinant $interferon-{\alpha}-A\;(rIFN-{\alpha}-A)$ on FVA infection. The extent of the infection was determined by measuring the weights of the spleens. Daily intraperitoneal injection of ddC (100 mg/kg body weight), $rIFN-{\alpha}-A$ (10 KU/mose) and the combination of both drugs to FVA inoculated mice for 18 days resulted in suppression of the growth of spleens by 15.1%, 52.7% and 61.6%, respectively. When ddC was dissolved in drinking water (0.1 mg/ml) and administered to a group of FVA inoculated mice ad libitum, and $rIFN-{\alpha}-A$ (10 KU/mouse) was intraperitoneally injected daily to another group of ddC (0.1 mg/ml) drinking mice for 18days, the growth of spleens was suppressed by 38.4% and 83.2%, respectively. These results indicate that administration of ddC via drinking water is more effective in suppressing FVA infection than the daily injection of ddC, and that the combined effects ddC and $rIFN-{\alpha}-A$ are not synergistic but additive. In order to determine whether ddC treatment alters the characteristic of the progenitor cells with respect to $Ca^{++}$ uptake, $Ca^{++}$ uptake in erythroid cells and the effect of cyclohexyladenosine (CHA) on the $Ca^{++}$ uptake were studied. $Ca^{++}$ uptake in the erythroid progenitor cells was about 20-fold greater than in mouse erythrocytes and the inhibition of $Ca^{++}$ uptake by CHA was the greatest in the progenitor cells from FVA infected mice which were treated with ddC. The inhibition was obviated by theophylline. Results of CHA binding studies showed that the erythroid progenitor cells contain both high and low affinity CHA binding sites, whereas mose erythrocytes contain only the low affinity CHA binding sites.

• Proceedings of the Korean Society of Toxicology Conference
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• 2003.10b
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• pp.87-88
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• 2003

PegIntron is the pegylated form of human recombinant interferon alfa-2b (IFN${\alpha}$2b). IFN ${\alpha}$2b, known as Intron A, has been in approved clinical use since the 1980's for various cancer indications, and for the treatment of Hepatitis C. In the mid 1990's, several clinical investigators reported that combination therapy with ribavirin and Intron A dramatically increased the therapeutic efficacy for treatment of Hepatitis C.(omitted)